IP Library › Granted Patent US 12,606,843
Granted Patent B2
US 12,606,843 · App. 16/927,679 · Granted Apr 21, 2026

CAS9 fusion proteins, RNA-guided gene editing, gene regulation compositions, and methods comprising the same

Inventors: Charles A. Gersbach (Durham, NC); Isaac B. Hilton (Durham, NC); Pablo Perez-Pinera (Urbana, IL); Ami M. Kabadi (Greensboro, NC); Pratiksha I. Thakore (Durham, IL); David G. Ousterout (Raleigh, NC); Joshua B. Black (Durham, NC)
Assignee: Duke University
C12N15/85A61K38/465A61K48/0058C07K14/4708C12N9/22C12N9/96C12N15/907A61K48/005C07K2319/71C12N2740/16043C12N2800/40C12N2840/20C12Y301/00
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Quick Facts
Patent No.
US 12,606,843
App. No.
16/927,679
Granted
Apr 21, 2026
Kind
B2
Abstract

Disclosed herein are Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas) 9-based system related compositions and methods of using said CRISPR/Cas9-based system related compositions for altering gene expression and genome engineering. Also disclosed herein are compositions and methods of using said compositions for altering gene expression and genome engineering in muscle, such as skeletal muscle and cardiac muscle.

Claims (15)

1 . A fusion protein comprising two heterologous polypeptide domains, wherein the first polypeptide domain comprises a Clustered Regularly Interspaced Short Palindromic Repeats associated (Cas) protein comprising a Cas9 derived from Streptococcus pyogenes and the mutations D10A and H840A to inactivate the nuclease activity of Cas9, and wherein the second polypeptide domain comprises a p300 histone acetyltransferase effector domain comprising a wild-type human p300 protein or the core lysine-acetyltransferase domain of the human p300 protein.

2 . The fusion protein of claim 1 , wherein the Cas protein comprises amino acids 36-1403 of SEQ ID NO: 1.

3 . The fusion protein of claim 1 , wherein the p300 histone acetyltransferase effector domain comprises a wild-type human p300 protein.

4 . The fusion protein of claim 1 , wherein the p300 histone acetyltransferase effector domain comprises the core lysine-acetyltranserase domain of the human p300 protein.

5 . The fusion protein of claim 1 , wherein the fusion protein activates transcription of a target gene by activating regulatory elements.

6 . The fusion protein of claim 5 , wherein the regulatory elements comprise a promoter of the target gene.

7 . The fusion protein of claim 5 , wherein the regulatory elements comprise an enhancer of the target gene.

8 . The fusion protein of claim 1 , wherein the fusion protein further comprises a linker connecting the first polypeptide domain to the second polypeptide domain.

9 . A kit comprising the fusion protein of claim 1 .

10 . An isolated polynucleotide encoding the fusion protein of claim 1 .

11 . A vector comprising the isolated polynucleotide of claim 10 .

12 . An isolated cell comprising the isolated polynucleotide of claim 10 .

13 . The fusion protein of claim 1 , wherein the second polypeptide domain comprises amino acids 1422-2038 of SEQ ID NO: 676.

14 . The fusion protein of claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 676.

15 . A method of modulating mammalian gene expression in a cell, the method comprises contacting the cell with the fusion protein of claim 1 .

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 13, 2021
From: GERSBACH, CHARLES A.; HILTON, ISAAC B.; PEREZ-PINERA, PABLO; KABADI, AMI M.; THAKORE, PRATIKSHA I.; OUSTEROUT, DAVID G.; BLACK, JOSHUA B.
To: DUKE UNIVERSITY
Reel/Frame 055922/0670 →
Continuity (8)
Continuation 15991333 · May 29, 2018
Division 14895316
Provisional Application 61981575 · Apr 18, 2014
Provisional Application 61967466 · Mar 19, 2014
Provisional Application 61904911 · Nov 15, 2013
Provisional Application 61839127 · Jun 25, 2013
Provisional Application 61831481 · Jun 5, 2013
Related Publication 20210032654A1 · Feb 4, 2021
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